Cosmetic preparation with a peptide addition

ABSTRACT

A cosmetic preparation contains peptide derivatives from α-MSH, as well as other active components. A cosmetic product has the property of activating the melanogenesis and being an anti-inflammatory and acting more efficiently. The synergetically active preparation also includes a combination of a peptide derivative corresponding to the formula (Lip)X-His-Phe-Arg-Y in a ratio of 0.05 mg to 2.5 mg of a pure peptide derivative per kg of the total mass, while the peptide derivative is mixed with xanthine in a ratio of 0.5 to 2 mol per 100 mol of peptide. Also there is at least 0.5 wt % of a mixture of enzymes and vitamins, containing at least 150 U/ml of superoxide dismutase. Also present are auxiliary agents and carrier agents in a ratio of 65 to 99.5 wt %, and possibly other active components.

BACKGROUND OF THE INVENTION

1. Field of the Invention

This invention concerns a new cosmetic preparation containing peptidederivatives derived from α-MSH (melanocyte stimulating hormone) andother active ingredients.

2. The Prior Art

α-MSH has already been investigated by a number of research groups,without so far being able to develop a pharmaceutical drug from it. Aspecific direction of effect was disclosed in French Patent 2,710,340 A,where certain peptide derivatives derived from α-MSH were described andclaimed as active ingredients which stimulate melanogenesis. Topical useagainst inflammatory reactions of the skin is also described.

SUMMARY OF THE INVENTION

The object of this invention is to develop a new cosmetic preparationespecially with melanogenesis-stimulating and anti-inflammatoryproperties with improved effectiveness.

According to this invention, a cosmetic preparation with a peptideadditive consists of a combination of the following active ingredients:

a) a peptide derivative of the formula [Lip]X-His-Phe-Arg-Y, where

Lip stands for thioctic acid or one of its derivatives,

X denotes Glu, OH, or NH₂,

Y is Trp-Gly-OH,

 Trp-Gly-NH₂,

 Trp-NH₂ or

 Trp-OH,

Phe is homo-Phe or P-fluoro-Phe,

and the amino acids may be in the form D, L or DL, or mixtures thereof,in a ratio of 0.05 to 2.5 mg pure peptide derivative per kg totalweight, where the peptide derivative is associated with xanthine in aratio of 0.5 to 2 mol per 100 mol peptide;

b) at least 0.5 wt % of a mixture of enzymes and vitamins containing atleast 150 units/mL (U/mL) peroxide dismutase;

c) the usual excipients and vehicles in the amount of 65 to 99.5 wt %,and

d) and additional active ingredients in the amount of 0 to 12 wt %.

The percentage amounts are based on the total weight of the cosmeticpreparation in each case.

A lipoyl peptide or peptide mixture known from French Patent No.2,710,340 A was used as the peptide; it has in particular at least oneof the following sequences:

I. [(DL)Lip]-Glu-His-D.HomoPhe-Arg-Trp-Gly-NH₂

II. [(DH)Lip]-Glu-His-D.HomoPhe-Arg-Trp-Gly-NH₂

III. [(DL)Lip]-Glu-His-parafluoro-Phe-Arg-Trp-Gly-NH₂

IV. [(DH)Lip]-His-D.HomoPhe-Arg-Trp-Gly-NH₂

V. [N.Lipoyl-lysine]-Glu-His-D.HomoPhe-Arg-Trp-Gly-NH₂

VI. [N.Lipoyl-lysine]-His-D.HomoPhe-Arg-Trp-Gly-NH₂

VII. [N.Lipoyl-lysine]-His-D.HomoPhe-Arg-Trp-NH₂

as well as derivatives of these molecules in the form of salts of theesters or amides, wherein the peptide derivative is mixed with xanthinein a ratio of 0.5 to 2 mol per 100 mol peptide.

In commercial peptides, e.g., MAP® or MAP-X® (from Laboratories Seporga,France), a lipoylaminopeptide of the above peptide derivative, the purepeptide content is about 50 mg/kg, and approximately 0.01 to 5 wt % ofthis is used in the cosmetic preparation. This yields theabove-mentioned peptide content for this invention.

The mixture of enzymes and vitamins used is preferably the digestionproduct of a yeast prepared by an ultrasonic treatment, with thedigestion product containing peroxide dismutase, protease, vitamin B₂,vitamin B₆, vitamin B₁₂, vitamin D₂, and vitamin E. It preferablycontains at least 150 U/mL peroxide dismutase (POD), protease, andvitamins B and D, with the POD/protease radio in international unitsbeing at least in the range of 3:1 to 8:1.

An ultrasonic digestion method, such as that described in German PatentNo. 4,241,154 C1 is especially advantageous for preparation of theenzyme/vitamin mixture according to this invention; in this process, acell dispersion or suspension is passed through an ultrasonic treatmentarea in an ultrasonic flow-through cell, with the sonotrode extendinginto the flow-through cell by one-half to two-thirds of its length sothat it is submerged in the medium to be treated ultrasonically. Thesonotrode here has an angle of 80.5° to 88.5°, and the ratio of thesubmersion length of the sonotrode in mm to the ultrasonically treatedvolume in mL is adjusted to a value of 1:1.1 to 1:20. The solids contentin the medium to be ultrasonically treated is in the range of 1:0.02 to1:2.2 (in wt %).

Yeasts, such as baker's yeast, brewer's yeast, wine yeast, as well asspecially treated yeasts such as POD-enriched yeasts, for example, canbe used as the cell dispersion. A cell dispersion that is advantageousto use may contain Saccharomyces cerevisiae, for example.

A particularly advantageous component a) of the cosmetic preparationaccording to this invention consists of a mixture of the peptidederivative of the following formula, mixed with xanthine:

[Lip]X-His-Phe-Arg-Y

manufactured with semisynthetic marine peptides and polypeptides whichare a bioengineered protein fraction produced from microalgae of theChlorella genus and with macroalgae of the Ulva genus associated withbyssus (silk filament from mollusks) and subsequently associated with avegetable glucose polymer, and where the semisynthetic marine peptidesare associated with 0.5 to 5 wt % marine mineral salts and traceelements.

An example of such a mixture is the product available under the brandname “Sun Marine Complex” (from Laboratories Seproge, Sophia-AntipolisCedex, France).

This mixture, which is advantageous to use, contains xanthine-associatedpeptides and semisynthetic marine peptides, the latter associated withglucose polymers, e.g., dextrin, and mineral salts/trace elements fromsea water, hereinafter referred to designated SMC (Sun Marine Complex);it is produced by simply mixing the two ingredients together. Thesemisynthetic marine peptides are produced by enzymatic purification ofthe tyrosine-rich peptides and polypeptides from byssus, i.e., from themarine mollusks. Then the vegetable cellulose component (microalgae andmacroalgae) is treated enzymatically to extract the various mineral andorganic elements. Then, under slightly basic conditions and under arelatively high pressure and high temperature, the association with aglucose polymer is induced, e.g., with dextrin to achieve stabilizationof the individual elements of the mixture on the gludosidic carrier as a“transport molecule” for the organism.

In SMC the ratio of xanthine-associated peptides is in the range of 0.5to 10 wt %, preferably 0.5 to 5 wt %. The remainder consists of thesemisynthetic marine peptides (associated with dextrin, for example)plus mineral salts and trace elements.

As the conventional additives and vehicles of the topical preparationaccording to this invention, the following can be used, e.g., isopropylmyristate, isopropyl palmitate, isopropyl stearate, carbomer, cetearylalcohol, lecithin, copolymers, paraffin oil, cetyl alcohol, propyleneglycol, polyglycol, jojoba oil, silicone oil, coconut oil, kaolinmodified according to WO 95/17,157, cetyl palmitate, C10-C30 alkylacrylate cross polymer, magnesium aluminum silicate,hydroxyethylcellulose, as well as other substances suitable for thespecific use forms, such as lipstick, eye cosmetics, hair mask products,etc., with which those skilled in the art are familiar.

The preparation may contain as additional active ingredients 1,3- and1,6-β-glucan, CM-Glucan®, allantoin, TiO₂, ZnO and UVA and UVB-blockingsubstances.

New agglomerates of nonporous spherical SiO₂ particles of 0.05-1.5 μmand spherical TiO₂ or ZnO particles in which the agglomerates have aparticle size of 0.06-5 μm can also be used in the emulsion in theamount of 0.1-30 wt %. Such agglomerates are produced by mixing theparticles while stirring at 300-400 rpm and adding some water untilachieving a pasty consistency and then adding the remaining water andhomogenizing at 3000 to 5000 rpm for 20-60 minutes.

The cosmetic preparation according to this invention surprisingly has asynergistic anti-inflammatory action which goes far beyond the potentialof the individual ingredients, and therefore it can be used successfullyin cosmetic compositions such as sunscreen emulsions, after-sunemulsions, facial cosmetics, etc., and as cosmetics for otherinflammatory dermatological processes. This synergism is demonstratedespecially impressively in the fact that an emulsion which contains only0.5 wt % of the xanthine-associated peptide preparation (with 50 mgpeptide per kg) plus the yeast digestion products has the sameanti-inflammatory effect as a pure peptide preparation (with 50 mgpeptide per kg).

This is shown by measurements on subjects with a Mexameter MX16®(Courage+Khazaka, Germany), using the degree of redness of the skin asthe basis for evaluation, with measurements being performed at certainintervals after the radiation exposure. The resulting curves as well asthe histological skin tests performed on the volunteers in parallel bythe H+E stain method show that with regard to melanocyte activity, thatthe activity of the preparation according to this invention is far abovethe activity levels to be expected of the individual ingredients.

A preferred formulation contains the peptide components in a ratio of0.05 to 1.5 mg pure peptide derivative per kg total weight.

Component a) according to this invention in the combination ofxanthine-associated peptides, peptide derivatives and the bioengineeredprotein fraction of enzyme activators containing semisynthetic marinepeptides—the latter are especially rich in tyrosine andphenylalanine—have a stimulating, protective, and regenerative effect onthe skin.

The skin is stimulated through the metabolism taking place with theinvolvement of AMPc, which is activated by peptides and xanthine. Thepeptide derivatives, especially MAPX®, stimulate melanin synthesis. Thesemisynthetic peptides regenerate the connective tissue.

The protective and regenerative effect consists first of the lightprotective effect caused by newly formed melanin, which plays the roleof a natural UV filter. It also consists of regeneration of UV-damagedcells by modulation of the cytokinins IL-1a and TNFα, as well as asynergistic effect of all the peptides present in the preparation withrespect to free radicals. The anti-inflammatory effect (anti-IL-1a andanti TNFα) was determined experimentally as 55% or 40%.

The yeast extract used produces, among other things, a better structureof the skin cells and an improved moisture balance.

Dermatological testing of skin to which an advantageous composition ofxanthine-associated peptides and semisynthetic marine peptides incombination with glucose polymers and marine mineral salts/traceelements had been applied showed an improvement in skin morphology tothe extent that the keratin was very well hydrated, and there were farfewer vacuoles and less edema in comparison with untreated sections ofskin. The dermis was well preserved and had normal connective tissue.The physical and chemical sunscreen filter content caused less skinirritation after being used for twenty-four hours.

The cosmetic preparation according to this invention can be used, forexample, in sun creams, sun gels, after-sun products, day creams, nightcreams, masks, body lotions, cleansing milk, make-up, lipstick, bodypowder, eye cosmetics, hair masks, hair rinse, hair shampoo, showergels, shower oils, bath oils. Such products are produced by methods withwhich those skilled in the art in this field are familiar.

This invention is described in greater detail below. The percentageamounts given in the examples are percent by weight (wt %).

BRIEF DESCRIPTION OF THE DRAWING

In the accompanying drawing,

FIG. 1 shows a diagram of the Mexameter measurement of reddened skinwith a comparison of various emulsions.

DETAILED DESCRIPTION OF PREFERRED EMBODIMENT Example 1

1A) Preparation of the yeast digestion product

A yeast suspension was prepared from yeast of the strain Saccharomycescerevisiae enriched with peroxide dismutase and the followingingredients:

20% yeast

10% propylene glycol

0.4% preservative

water to 100%.

The ingredients were mixed and cooled to about 8 to 10° C. The yeastsuspension was treated according to Example 1 of German Patent No.4,241,154 C1, with approximately three liters of digestion product beingremoved per hour from the flow-through cell at a maximum temperature of20° C. After separating the cell walls, a product with the followingactive ingredient contents is obtained:

peroxide dismutase ≧200 U/mL protease ˜50 U/mL (U = units) vitamin B₂ 20mg/L vitamin E 0.6 mg/mL vitamin B₆ 40 mg/L vitamin B₁₂ 3 mg/L vitaminD₂ 0.3 mg/mL

1B) Preparation of the cosmetic emulsion

The following examples were conducted with the yeast digestion productaccording to Example 1A and with the following general procedure inExamples 2-4.

Phase B, heated to about 80° C., was added to phase A at approximately80° C. while stirring. The mixture was homogenized, cooled and mixedwith phase C at about 35° C. The mixture was homogenized.

Example 2 Sun Cream

Preparation according to Example 1B

Phase A glycerin 3.0% magnesium sulfate 0.5% water to 100% Phase Bglyceryl oleate 2.5% decyl oleate 5.0% paraffin oil 10.0% beeswax 2.0%TiO₂ 3.0% zinc stearate 2.0% Phase C preservative 0.3% perfume oil 0.5%peptide preparation MAP-X ® 1.0% yeast digestion product according to0.5%

Example 3 After-sun Preparation

Product prepared according to Example 1B

Phase A Glycerin 6.0% Magnesium sulfate 1.0% Water to 100% Phase BGlyceryl oleate 4.0% Polyglyceryl-3 diisostearate 10.0% Hexyldecanol2.0% Beeswax 1.0% Dicapryl ether 2.0% Phase C Preservative 0.4% Perfumeoil 0.5% Peptide preparation MAP-X ® 2.0% Yeast digestion productaccording to 5.0%

Example 4 Body Lotion

Lotion prepared as in Example 1B

Phase A Glycerin 3.0% Magnesium sulfate 0.5% Propylene glycol 2.0% Waterto 100% Phase B Glyceryl oleate 1.0% Polyglyceryl-2dipolyhydroxystearate 2.5% Cetearyl ionanoate 3.0% Jojoba oil 1.5% PhaseC Preservative 0.4% Perfume oil 1.5% Peptide preparation MAP-X ® 3.0%Yeast digestion product according to 8.0%

Example 5 Lipstick

Phase A was melted at about 85° C. Phase B was stirred well with thecolor, and phase A was added. Then phase C was added and stirred, andthe mixture was cooled.

Phase A Candelilla wax (vegetable wax) 8.0% Beeswax to 100% Phase BCeraphyl oil 25.0% Calendula oil 25.0% Pigment (depending on color) 8.0%Phase C Peptide preparation MAP-X ® 2.0% Yeast digestion productaccording to 2.0% Fragrance 1.0%

Example 6 Shower Gel

Water was placed in the vessel first, and then the following ingredientswere added in the usual manner while stirring and then homogenizing.

Water to 100% Disodium myreth sulfate 25% Disodium laurethsulfosuccinate 8% Preservative 0.2% Pefume oil 0.5% Peptide preparationMAP-X ® 0.5% Yeast digestion product according to 0.1%

Example 7

FIG. 1 shows a comparison of different emulsions in measurement of thereddening of the skin of volunteers. The values are the averages of tenmeasurements. The initial value 1 was arbitrarily set as a dimensionlessproportionality factor (index). The measurements are performed at thetimes indicated in the curves. The measurements were made using aMexameter® MX16 from the company Courage+Khazaka Electronic GmbH,Germany. Absorption of wavelengths 568 and 660 nm is measured here, withone wavelength corresponding approximately to the absorption peaks ofhemoglobin in the skin, and the other wavelength preferably excludingother color influences (e.g., bilirubin) as much as possible. Themeasured values thus obtained are given as the proportionality factor(index) of the colors used.

The following were compared:

A a base emulsion containing only the emulsion base and 0.5 wt % of theyeast digestion product according to Example 1A (“base”)

B peptide preparation MAP-X® in a concentration of 50 mg pure peptideper kg (“peptide”)

C same base emulsion as in A with 0.5% peptide preparation MAP-X®(“peptide+base”).

The shape of the curve after 24 hours and 48 hours shows clearly thatemulsion C has an unexpected improvement in relation to A or B.

Since the effect of antioxidants must be evaluated in the manner ofcomplex physiological control circuits in a biological system, a simpleadditive effect of the two substances with the given antioxidant effectis not observed. Instead, an overall effect only slightly higher thanthe normal value is to be expected. Thus the difference in measurementresults must clearly be regarded as a synergistic effect.

Example 8 24-hour Skin Protection Cream (Before-sun and After-sunProtection)

Phase A C12-15 alkylbenzoate 4.0% Shea butter 2.0% Steareth-2 1.5% PhaseB Distilled water to 100% Cross polymer 0.5% Glycerin 2% Phase CTriethanolamine 0.5% Phase D Jojoba oil 2% olive oil 1% Preservative0.5% Phase E Sun Marine Complex 5.0% Yeast digestion product accordingto 0.5% Perfume oil 0.2%

The cream was prepared by stirring the ingredients of phases A and Bseparately at about 60° C., then mixing the two phases together at thistemperature, and finally cooling to about 45° C. Next, phases C and Dwere added and the mixture was cooled while stirring. Phase E was addedat 35° C. and stirred with the overall mixture.

Example 9 Sun Cream (SPF 10)

Phase A Steareth-2 3% Steareth-21 2% Beeswax 1.5% Phase B Distilledwater to 100% Glycerin 3.5% Propylene glycol 2.0% TiO₂ 7.0% Phase CJojoba oil 2.5% Babassu oil 1.0% Silicone oil 0.5% Preservative 0.3%Phase D Sun Marine Complex 0.5% Yeast digestion product according to0.5% Perfume oil 0.1%

It is prepared as described in Example 8.

Example 10 Sun Cream (SPF 15)

Phase A Steareth-2 2.0% Steareth-21 2.0% Isohexadecane 3.0% Octylmethoxycinnamate 5.0% 4-methylbenzylidene camphor 3.3% Phase B Distilledwater to 100% Glycerin 10.0% TiO₂/SiO₂ agglomerate* 2.0% ZnO/SiO₂agglomerate* 1.0% Phase C Silicone oil 2.0% Palm oil 4.0% Preservative0.3% Phase D Sun Marine Complex 0.5% Yeast digestion product accordingto 0.5% Perfume oil 0.2% *from spherical nonporous SiO₂ particles of0.05-1.5 μm and spherical TiO₂ and ZnO particles, where the agglomerateshave a particle size of 0.06-1.5 μm.

The preparation was processed according to Example 8.

Example 11 Sun Cream with Chemical Filters

Phase A Cetearyl 1.5% Glyceryl steareth, ceterareth 20, cetyl 3.5% Octylsteareth 1.5% OctYl methoxycinnamate 6.5% 4-Methylbenzylidene camphor1.5% Phase B Distilled water to 100% Glycerin 2.0% Phase C Babassu oil5.0% Preservative 0.5% Phase D Sun Marine Complex 3.0% Yeast digestionproduct according to 0.5% Perfume oil 0.1%

Cream processed according to Example 8.

Example 12 Make-up with SPF 4

Phase A Shea butter 2% Beeswax 3% olive oil 5% Color 3-10% TiO₂ 4% PhaseB Distilled water to 100% Glycerin 2% Phase C Jojoba oil 2% Silicone oil5%

The composition was processed according to Example 8.

What is claimed is:
 1. A cosmetic preparation with a peptide additive,comprising as active ingredients a combination of the followingingredients: a) a peptide derivative of the formula(Lip)X-His-Phe-Arg-Y, where Lip represents thioctic acid or one of itsderivatives, X denotes Glu, OH, or NH₂ Y denotes Trp-Gly-OH,Trp-Gly-NH₂, Trp-NH₂ or Phe denotes Homo-Phe or P-fluoro-Phe, and theamino acids may be present in the form D, L or DL, or mixtures thereof,in the amount of 0.05 to 2.5 mg pure peptide derivative per kg totalweight, where the peptide derivative is mixed with xanthine in a ratioof 0.5 to 2 mol per 100 mol peptide; b) at least 0.5 wt % of a mixtureof enzymes and vitamins containing at least 150 U/mL peroxide dismutase(POD); c) 65 to 99.5 wt % of an inert, non-toxic cosmetically acceptablecarrier; and d) 0 to 12 wt % other active ingredients, where thepercentage amounts are all based on the total weight of the preparation.2. A preparation according to claim 1, wherein the mixture of enzymesand vitamins, of (b) are peroxide dismutase, protease, vitamin B2,vitamin B6, vitamin B12, and vitamin E.
 3. A preparation according toclaim 1, wherein it contains protease and vitamins B and D, with theratio of peroxide dismutase to protease, expressed as internationalunits, being in the range of 3:1 to 8:1.
 4. A preparation according toclaim 2, wherein the mixture of enzymes and vitamins of (b) originatesfrom ultrasonic digestion of a yeast.
 5. A preparation according toclaim 1, wherein the peptide derivative of (a) is selected from thegroup consisting of I ((DL)Lip)-Glu-His-D.HomoPhe-Arg-Trp-Gly-NH₂ II((DH)Lip)-Glu-His-D.HomoPhe-Arg-Trp-Gly-NH₂ III((DL)Lip)-Glu-His-para-fluoro-Phe-Arg-Trp-Gly-NH₂ IV((DH)(Lip)-His-D.HomoPhe-Arg-Trp-Gly-NH₂ V(N.Lipoyl-lysine)-Glu-His-D.HomoPhe-Arg-Trp-Gly-NH₂ VI(N.Lipoyl-lysine)-His-D.HomoPhe-Arg-Trp-Gly-NH₂ VII(N.Lipoyl-lysine)-His-D.HomoPhe-Arg-Trp-NH₂ and as well as derivativesof these molecules in a form of salts of esters or amides.
 6. Apreparation according to claim 1, wherein the peptide component ispresent in the amount of 0.05 to 1.5 mg pure peptide derivative per kgtotal weight.
 7. A preparation according to claim 1, wherein the peptidederivative mixed with xanthine is present in the form of a mixture ofpeptide derivatives of the formula (Lip)X-His-Phe-Arg-Y and marinepeptides and polypeptides, which are a protein fraction produced frommicro-algae of the genus Chlorella and macroalgae of the genus Ulvamixed with byssus (mollusk silk), then mixed with a vegetable glucosepolymer, and wherein marine peptides are mixed with 0.5 to 5 wt % marinemineral salts and trace elements.
 8. A preparation according to claim 7,wherein it is a mixture of water, dextrin glycoproteins together with amixture of Ulva and Chlorella algae extract and mollusk silk.
 9. Acosmetic preparation with a peptide additive according to claim 1wherein the inert, non-toxic cosmetically acceptable carrier of (c) isselected from the group consisting of sun cream, sun emulsion, after-sunlotion, before-sun lotion, body lotion, lipstick, make-up, eye cosmetic,face powder, hair mask, hair shampoo, hair lotion, shower gel and showeroil.
 10. A preparation according to claim 4, wherein the mixture ofenzymes and vitamins of (b) originate from ultrasonic digestion ofbaker's yeast.